Handa K, Jacobs F, Longenecker B M, Hakomori S I
Division of Biomembrane Research, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA.
Biochem Biophys Res Commun. 2001 Jul 20;285(3):788-94. doi: 10.1006/bbrc.2001.5225.
Two mucin-type glycoproteins, MUC-1 and P-selectin glycoprotein ligand-1 (PSGL-1), and glycosphingolipids (GSLs), expressed in human T-cell line HUT78, were highly enriched in low-density buoyant fraction (termed "GEM"), together with CD45, Yes, Fyn, and lck(56). Enrichment of MUC-1, PSGL-1 and GSLs, together with these signal transducer molecules in low-density membrane fraction was observable when fraction was prepared from cells either in nonionic detergent Brij 58 or in hypertonic alkaline conditions (500 mM Na(2)CO(3)). On pretreatment of cells with cholesterol-binding reagent methyl beta-cyclodextrin, levels of MUC-1 and PSGL-1 together with the above signal transducers in GEM was greatly reduced, and they were translocated into high-density membrane fraction. Similar association of lck(56), Yes, Fyn, and cSrc together with MUC-1 was also found in GEM fraction of mouse T-cell lymphoma EL4 cells expressing MUC-1 through transfection of its gene. These findings indicate the presence of another glycosyl cluster ("glycocluster"), in addition to the previously well-established GSL cluster organized with signal transducer molecules in GEM fraction, and its possible functional role in T-cells.
在人T细胞系HUT78中表达的两种粘蛋白型糖蛋白,即MUC-1和P-选择素糖蛋白配体-1(PSGL-1)以及糖鞘脂(GSLs),与CD45、Yes、Fyn和lck(56)一起,在低密度漂浮组分(称为“GEM”)中高度富集。当用非离子去污剂Brij 58或在高渗碱性条件(500 mM Na₂CO₃)下从细胞制备组分时,可观察到MUC-1、PSGL-1和GSLs以及这些信号转导分子在低密度膜组分中的富集。在用胆固醇结合试剂甲基-β-环糊精预处理细胞后,GEM中MUC-1和PSGL-1以及上述信号转导分子的水平大大降低,并且它们转移到高密度膜组分中。在通过基因转染表达MUC-1的小鼠T细胞淋巴瘤EL4细胞的GEM组分中,也发现了lck(56)、Yes、Fyn和cSrc与MUC-1的类似关联。这些发现表明,除了先前在GEM组分中与信号转导分子一起建立的GSL簇之外,还存在另一种糖基簇(“糖簇”),及其在T细胞中的可能功能作用。